Product datasheet
Anti-SMC1 antibody ab21583
2 Abreviews 5 References 5 Images
Overview
Product name
Anti-SMC1 antibody
Description
Rabbit polyclonal to SMC1
Tested applications
ICC/IF, WB, IP, IHC-P
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Chicken, Xenopus laevis
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1200 to the C-terminus of
Human SMC1. Read Abcam's proprietary immunogen policy (Peptide available as ab23863.)
Positive control
HeLa and Jurkat whole cell lysate
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or 80°C. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab21583 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Notes
ICC/IF
Use a concentration of 1 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 150 kDa
(predicted molecular weight: 143 kDa).
IP
Use at an assay dependent concentration.
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Application
Abreviews
IHC-P
Notes
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before
commencing with IHC staining protocol.
Target
Anti-SMC1 antibody images
All lanes : Anti-SMC1 antibody (ab21583) at
1 µg/ml
Lane 1 : 20ug HeLa (Human epithelial
carcinoma cell line) Whole Cell Lysate
Lane 2 : Jurkat (Human) Whole Cell Lysate
(ab7899) at 20 µg
Lane 3 : 20ug HeLa (Human epithelial
carcinoma cell line) Whole Cell Lysate with
Human SMC1 peptide (ab23863) at 1 µg/ml
Lane 4 : Jurkat (Human) Whole Cell Lysate
Western blot - SMC1 antibody (ab21583)
(ab7899) at 20 µg with Human SMC1 peptide
(ab23863) at 1 µg/ml
Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor®
680) (ab28446) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 143 kDa
Observed band size : 150 kDa
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SMC1 was immunoprecipitated using 0.5mg
Hela whole cell extract, 5µg of Rabbit
polyclonal to SMC1 and 50µl of protein G
magnetic beads (+). No antibody was added
to the control (-).
The antibody was incubated under agitation
with Protein G beads for 10min, Hela whole
cell extract lysate diluted in RIPA buffer was
added to each sample and incubated for a
further 10min under agitation.
Proteins were eluted by addition of 40µl SDS
Immunoprecipitation - Anti-SMC1 antibody
loading buffer and incubated for 10min at
(ab21583)
70oC; 10µl of each sample was separated on
a SDS PAGE gel, transferred to a
nitrocellulose membrane, blocked with 5%
BSA and probed with ab21583.
Secondary: Goat polyclonal to mouse IgG
light chain specific (HRP) at 1/5000 dilution.
Band: 150kDa: SMC1; Non specific - 41 and
42kDa: We are unsure as to the identity of
this extra band.
ICC/IF image of ab21583 stained human
HeLa cells. The cells were methanol fixed (5
min) and incubated with the antibody
(ab21583, 1µg/ml) for 1h at room
temperature. The secondary antibody
(green) was Alexa Fluor® 488 goat anti-rabbit
Immunocytochemistry/ Immunofluorescence SMC1 antibody (ab21583)
IgG (H+L) used at a 1/1000 dilution for 1h.
Image-iTTM FX Signal Enhancer was used as
the primary blocking agent, 5% BSA (in TBST) was used for all other blocking steps. DAPI
was used to stain the cell nuclei (blue). Alexa
Fluor® 594 WGA was used to label plasma
membranes (red).
Panel A shows localisation of ab21583 to the
nuclei, Panel B has the Alexa Fluor® 488
channel removed for comparison.
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ab21583 staining SMC1 in assynchonous
HeLa cells (green). Cells were
paraformaldehyde-fixed (4% - 10min) and
counterstained with DAPI (red). Secondary
antibody: Goat anti-Rabbit conjugated to Cy3
®. Please refer to Abreview for further details.
Immunocytochemistry/ Immunofluorescence SMC1 antibody (ab21583)
This image is courtesy of an Abreview submitted by
Dr Kirk McManus
IHC image of SMC1 staining in human skin
FFPE section, performed on a Leica BondTM
system using the standard protocol F. The
section was pre-treated using heat mediated
antigen retrieval with sodium citrate buffer
(pH6, epitope retrieval solution 1) for 20 mins.
The section was then incubated with
ab21583, 1µg/ml, for 15 mins at room
temperature and detected using an HRP
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) - SMC1 antibody
(ab21583)
conjugated compact polymer system. DAB
was used as the chromogen. The section was
then counterstained with haematoxylin and
mounted with DPX.
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